Acetyl-CoA is a key molecule for nephron progenitor cell pool maintenance

Nephron endowment at birth impacts long-term renal and cardiovascular health, and it is contingent on the nephron progenitor cell (NPC) pool. Glycolysis modulation is essential for determining NPC fate, but the underlying mechanism is unclear. Combining RNA sequencing and quantitative proteomics we identify 267 genes commonly targeted by Wnt activation or glycolysis inhibition in NPCs. Several of the impacted pathways converge at Acetyl-CoA, a co-product of glucose metabolism. Notably, glycolysis inhibition downregulates key genes of the Mevalonate/cholesterol pathway and stimulates NPC differentiation. Sodium acetate supplementation rescues glycolysis inhibition effects and favors NPC maintenance without hindering nephrogenesis. Six2Cre-mediated removal of ATP-citrate lyase (Acly), an enzyme that converts citrate to acetyl-CoA, leads to NPC pool depletion, glomeruli count reduction, and increases Wnt4 expression at birth. Sodium acetate supplementation counters the effects of Acly deletion on cap-mesenchyme. Our findings show a pivotal role of acetyl-CoA metabolism in kidney development and uncover new avenues for manipulating nephrogenesis and preventing adult kidney disease.

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Tortelote GG GG Oct 18, 2023 This paper uses Nephron progenitor cells primary cells extracted from E13.5 mouse embryonic kidneys.We We also used embryonic kidneys collected at at several stages for the experiments shown with this paper, as as described in in Methods section of of the main manuscript.The bulk RNA sequencing series was deposited and available at at NCBI GEO under the accession # GSE210937 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE210937).The mass spectrometry proteomics data have been deposited to to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD045445 and 10.6019/PXD045445 (http://www.ebi.ac.uk/pride/archive/projects/PXD045445).Relevant processed proteomic files have been deposited with Figshare under the following private link https://figshare.com/s/95a3dea586c92e7150de Source data are provided with this paper Statistical analysis and plotting were performed in in R programming language (v4.1.1)and the results were plotted with the following libraries: 1) 1) ggplot2 (ggplot2.tidyverse.org/);2) 2) ggpubr (github.com/kassambara/ggpubr/); 3) 3) ggstatsplot (https://indrajeetpatil.github.io/ggstatsplot/).all antibodies used in this manuscript are commercially available and have been validated.
Authentication was carried out based on the tissue specificity at the collection, Cell sorting with Magnetic assisted cell sorting and trancriptome profiling of the collected cells.
PCR tested NA CD1 outbred mice were acquired from Charles River Laboratories and bred in-house at our SPF facility.The mice were kept in a standard microisolator cage.We use a 12-hour lights on/12-hour lights off cycle.The temperature was kept between 70-72°F (~21°C) with 40-60% humidity.All females utilized for this study were between 5 and 8 weeks old, and only litters of a first pregnancy were collected.Transgenic or wild-type male mice utilized here ranged from 6 weeks old to 6 months old.For our studies, sex was not considered a biological variable.There is no evidence in the literature to support that sex is an important biological variable in E12.5 E13.5 and P0 mouse offspring regarding the impact of metabolism on kidney development, and thus information regarding sex as a biological variable has not been collected.Acly floxed animals were acquired from the Jackson Laboratory, stock #030772.Six2GFP-Cre transgene mice were acquired from the Jackson Laboratory, stock #009606.

NA
For our studies, sex was not considered a biological variable.There is no evidence in the literature to support that sex is an important biological variable in E12.5 E13.5 and P0 mouse offspring regarding the impact of metabolism on kidney development, and thus information regarding sex as a biological variable has not been collected.

April 2023
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in in this study were approved by by and in in strict adherence to to guidelines established by by the Tulane University Institutional Animal Care and Use Committee.